scholarly journals Development and Evaluation of an Immunochromatographic Strip for Detection of Streptococcus Suis Type 2 Antibody

2007 ◽  
Vol 19 (4) ◽  
pp. 355-361 ◽  
Author(s):  
Junxing Yang ◽  
Meilin Jin ◽  
Jianfeng Chen ◽  
Ying Yang ◽  
Pei Zheng ◽  
...  
Keyword(s):  
Capsular Polysaccharide ◽  
Protein A ◽  
Streptococcus Suis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Staphylococcal Protein A ◽  
Immunochromatographic Strip ◽  
Specificity And Sensitivity ◽  
Antibody Titers ◽  
Serological Surveillance ◽  
And Control

In this study, an immunochromatographic strip (ICS) was developed for the detection of antibody against Streptococcus suis serotype 2 (SS2). Colloidal gold particles labeled with staphylococcal protein A (SPA), which can bind to the FC fragment of mammalian immunoglobulin, were used as the detector reagent. The capsular polysaccharide (CPS) of SS2 and affinity-purified IgG from a healthy naive pig were immobilized on test and control regions of a nitrocellulose membrane, respectively. The ICS was used to 1) detect anti-CPS antibody in 14 sera taken from 4 SS2-infected pigs, 24 sera from pigs hyperimmunized with SS2, and 68 sera from pigs inoculated or infected with bacteria other than SS2; 2) determine anti-CPS antibody titers of 20 positive sera for comparison with enzyme-linked immunosorbent assay (ELISA); and 3) detect anti-CPS antibody in 226 clinical sera taken from diseased pigs also for comparison with ELISA. An ELISA used as a reference test determined the specificity and sensitivity of the ICS to be 97.1% and 86.3%, respectively. There was excellent agreement between the results obtained by ELISA and the ICS (kappa = 0.843). Additionally, there was strong agreement between the results of bacterial isolation from pig tonsils and ICS test (kappa = 0.658). Because it is rapid and easy to use, the test is suitable for the serological surveillance of SS2 at farms.

scholarly journals Evaluation of a Novel Enzyme-Linked Immunosorbent Assay for Detection of Antibodies against Salmonella, Employing a Stable Coating of Lipopolysaccharide-Derived Antigens Covalently Attached to Polystyrene Microwells

2000 ◽  
Vol 12 (2) ◽  
pp. 130-135 ◽  
Author(s):  
Camilla Wiuff ◽  
Eva Stenbæk Jauho ◽  
Henrik Stryhn ◽  
Lars Ole Andresen ◽  
Karina Thaulov ◽  
...  
Keyword(s):  
Microtiter Plate ◽  
Enzyme Linked Immunosorbent Assay ◽  
O Antigen ◽  
Salmonella Infections ◽  
Specificity And Sensitivity ◽  
Conventional Elisa ◽  
Polysaccharide Antigens ◽  
Serological Surveillance ◽  
Coated Surface ◽  
High Degree

Polysaccharides derived from Salmonella typhimurium lipopolysaccharide (LPS) representing the O-antigen factors 1, 4, 5, and 12 and the O-antigen factors 6 and 7 from Salmonella choleraesuis LPS were derivatized with the photoreactive compound anthraquinone and subsequently covalently coupled to microtiter polystyrene plates by ultraviolet irradiation. Both polysaccharide antigens could be coupled simultaneously to the same microtiter plate. The coated surface was used in indirect ELISA for the determination of serum antibodies from pigs infected with bacteria of the two Salmonella groups and from uninfected pigs. This ELISA proved itself by having a good long-term durability and a high degree of reproducibility, including low day-to-day variations and low interplate variations. Furthermore, the ELISA showed good specificity and sensitivity when data were compared with the optical density levels of a panel of pig sera as determined by a conventional ELISA on the basis of passive coating of the two Salmonella LPS antigens (the mix-ELISA). The covalent anthraquinone mix-ELISA shows promise as a stable and durable alternative to the existing conventional ELISA for serological surveillance of Salmonella infections in pigs.

scholarly journals Analysis of Human Serum Immunoglobulin G against O-Acetyl-Positive and O-Acetyl-Negative Serogroup W135 Meningococcal Capsular Polysaccharide

2005 ◽  
Vol 12 (5) ◽  
pp. 586-592 ◽  
Author(s):  
Peter C. Giardina ◽  
Emma Longworth ◽  
Renee E. Evans-Johnson ◽  
Michaelene L. Bessette ◽  
Hong Zhang ◽  
...  
Keyword(s):  
Immunoglobulin G ◽  
Capsular Polysaccharide ◽  
Solid Phase ◽  
Geometric Mean ◽  
Serum Antibody ◽  
Fluid Phase ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antibody Titers ◽  
The Impact

ABSTRACT The capsular polysaccharide of Neisseria meningitidis serogroup W135 is expressed in both O-acetyl-positive (OA+) and O-acetyl-negative (OA−) forms. This study investigates the impact of OA status (OA+ versus OA−) on serological measurements of anti-W135 immunoglobulin G (IgG) antibodies in immunized adults. W135-specific serum antibody assignments were made for 28 postimmunization sera from adults by enzyme-linked immunosorbent assay using the meningococcal standard reference serum CDC1992. The established IgG concentration in micrograms per milliliter ([IgG]μg/ml) for CDC1992 against OA+ antigen (16.2 μg/ml) was used as a reference to assign a concentration of 10.13 μg/ml IgG against OA− antigen by cross-standardization. Overall, the IgG assignments for these sera were higher against OA+ antigen (geometric mean concentration [GMC] = 7.16 μg/ml) than against OA− antigen (GMC = 2.84 μg/ml). However, seven sera showed higher specific [IgG]μg/ml values against the OA+ antigen than against the OA− antigen. These sera were also distinguished by the inability of fluid-phase OA− antigen to compete for antibody binding to OA+ solid-phase antigen. Although there was no overall difference in functional activity measured by complement-mediated serum bactericidal assay (SBA) against OA+ and OA− target bacteria (geometric mean titers of 9,642 and 9,045, respectively), three serum specimens showed a large difference in SBA antibody titers against OA+ versus OA− W135 target bacteria, which may reflect different epitope specificities for these sera. Our data indicate that, for some sera, the agreement in anti-OA+ versus anti-OA− W135 IgG assignments is serum specific and does not reflect the functional (killing) activity in vitro.

scholarly journals Identification of glycoprotein Ib as a target for autoantibody in idiopathic (autoimmune) thrombocytopenic purpura

Blood ◽  
1986 ◽  
Vol 67 (2) ◽  
pp. 310-315 ◽  
Author(s):  
NS Szatkowski ◽  
TJ Kunicki ◽  
RH Aster
Keyword(s):  
Direct Evidence ◽  
Protein A ◽  
Normal Subjects ◽  
Platelet Membrane ◽  
Staphylococcal Protein A ◽  
Glycoprotein Ib ◽  
Thrombocytopenic Purpura ◽  
One Dimensional ◽  
Rocket Electrophoresis ◽  
And Control

Abstract An antibody (DIL) from a patient with idiopathic thrombocytopenic purpura (ITP) was shown to have autospecificity on the basis of reactions with autologous platelets that were identical to those obtained with platelets from normal subjects. DIL antibody also reacted strongly in an immunofluorescence test with platelets from a patient with Glanzmann's thrombasthenia, but failed to react with platelets from a patient with the Bernard-Soulier syndrome who was known to be deficient in glycoprotein Ib (GPIb). Purified GPIb and control platelets, but not Bernard-Soulier platelets, inhibited the lytic activity of DIL. Using the GPIb-specific monoclonal antibody AP1 and one-dimensional rocket electrophoresis into gels containing rabbit antihuman platelet membrane antibody, it was shown that staphylococcal protein A-Sepharose beads coated with DIL antibody selectively remove GPIb from solubilized platelet preparations. By crossed immunoelectrophoresis it was found that DIL recognizes a determinant on GPIb on the membrane side of the cleavage site of the platelet calcium- activated protease (calpain). These studies provide direct evidence for binding of a platelet autoantibody to a determinant on GPIb relatively close to the site of insertion of this protein into the platelet membrane.

scholarly journals Identification of immunoreactive sites in bovine parathyroid cells to antibodies raised against the NH2-terminal sequence of parathyroid hormone.

1979 ◽  
Vol 27 (8) ◽  
pp. 1209-1214 ◽  
Author(s):  
W Limacher ◽  
P Wild ◽  
E Manser ◽  
H Lutz
Keyword(s):  
Parathyroid Hormone ◽  
Protein A ◽  
Electron Microscopic Level ◽  
Enzyme Linked Immunosorbent Assay ◽  
Staphylococcal Protein A ◽  
Terminal Sequence ◽  
Electron Microscopic ◽  
Secretion Granules ◽  
Parenchymal Cells ◽  
Bovine Parathyroid Hormone

The NH2-terminal sequence of bovine parathyroid hormone (1-84) was localized with different immunocytochemical methods on the light and electron microscopic level in bovine parathyroid glands and in isolated bovine parathyroid parenchymal cells. The peroxidase labeled staphylococcal protein A and the peroxidase anti-peroxidase method were found to be advantageous for light and electron microscopic localization, respectively. Reaction product was found light microscopically in the cytoplasma of the parenchymal cells and electron microscopically largely over the secretion granules of the parenchymal cells. The immunoreactive sites were subsequently identified to represent only intact parathyroid hormone (1-84) by gel electrophoresis derived enzyme linked immunosorbent assay.

scholarly journals Comparison of techniques for demonstrating antibodies to Rift Valley fever virus

1986 ◽  
Vol 97 (2) ◽  
pp. 317-329 ◽  
Author(s):  
R. Swanepoel ◽  
J. K. Struthers ◽  
M. J. Erasmus ◽  
S. P. Shepherd ◽  
G. M. McGillivray ◽  
...  
Keyword(s):  
Rift Valley ◽  
Rift Valley Fever ◽  
Rift Valley Fever Virus ◽  
Protein A ◽  
Haemagglutination Inhibition ◽  
Enzyme Linked Immunosorbent Assay ◽  
Staphylococcal Protein A ◽  
Valley Fever ◽  
Neutralization Techniques ◽  
Comparison Of Techniques

SummaryNine serological techniques were compared by monitoring the response to infection with Rift Valley fever (RVF) virus in three sheep. Antibodies were monitored daily for the first 14 days after infection, then weekly and later fortnightly up to week 24. The earliest antibody response was detected in one sheep on day 3 by a plaque reduction neutralization test, and by day 6 antibodies were demonstrable in all three sheep by haemagglutination-inhibition, reversed passive haemagglutination-inhibition, immunodiffusion, indirect immunofluorescence (IF), enzyme-linked immunosorbent assay and neutralization of cytopathic effect in cell cultures. Antibodies were demonstrable by complement fixation on day 8 at the earliest. IF and the two neutralization techniques produced the highest titres, but all tests could be used satisfactorily for the serological diagnosis of RVF. Inactivated antigen could be used for all except the neutralization tests. A radioimmunoassay technique using125I-labelIed staphylococcal protein A detected antibodies on day 8 at the earliest and produced lower mean titres than some of the other techniques. This was probably because sheep immunoglobulins bind protein A poorly.

scholarly journals Impaired Function of Antibodies to Pneumococcal Surface Protein A but Not to Capsular Polysaccharide in Mexican American Adults with Type 2 Diabetes Mellitus

2012 ◽  
Vol 19 (9) ◽  
pp. 1360-1369 ◽  
Author(s):  
Christine E. Mathews ◽  
Eric L. Brown ◽  
Perla J. Martinez ◽  
Upasana Bagaria ◽  
Moon H. Nahm ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Type 2 Diabetes ◽  
Type 2 Diabetes Mellitus ◽  
Capsular Polysaccharide ◽  
Protein A ◽  
Surface Protein ◽  
Complement C3 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Complement Deposition

ABSTRACTThe goal of the study was to determine baseline protective titers of antibodies toStreptococcus pneumoniaesurface protein A (PspA) and capsular polysaccharide in individuals with and individuals without type 2 diabetes mellitus. A total of 561 individuals (131 individuals with diabetes and 491 without) were screened for antibodies to PspA using a standard enzyme-linked immunosorbent assay (ELISA). A subset of participants with antibodies to PspA were retested using a WHO ELISA to determine titers of antibodies to capsular polysaccharide (CPS) (serotypes 4, 6B, 9V, 14, 18C, 19A, 19F, and 23F). Functional activity of antibodies was measured by assessing their ability to enhance complement (C3) deposition on pneumococci and promote killing of opsonized pneumococci. Titers of antibodies to protein antigens (PspA) were significantly lower in individuals with diabetes than controls without diabetes (P= 0.01), and antibodies showed a significantly reduced complement deposition ability (P= 0.02). Both antibody titers and complement deposition were negatively associated with hyperglycemia. Conversely, titers of antibodies to capsular polysaccharides were either comparable between the two groups or were significantly higher in individuals with diabetes, as was observed for CPS 14 (P= 0.05). The plasma specimens from individuals with diabetes also demonstrated a higher opsonophagocytic index against CPS serotype 14. Although we demonstrate comparable protective titers of antibodies to CPS in individuals with and individuals without diabetes, those with diabetes had lower PspA titers and poor opsonic activity strongly associated with hyperglycemia. These results suggest a link between diabetes and impairment of antibody response.

scholarly journals Vancomycin-dependent antibodies associated with thrombocytopenia and refractoriness to platelet transfusion in patients with leukemia

Blood ◽  
1990 ◽  
Vol 75 (2) ◽  
pp. 518-523 ◽  
Author(s):  
DJ Christie ◽  
N van Buren ◽  
SS Lennon ◽  
JL Putnam
Keyword(s):  
Platelet Transfusion ◽  
Protein A ◽  
Enzyme Linked Immunosorbent Assay ◽  
Severe Thrombocytopenia ◽  
Staphylococcal Protein A ◽  
Igg Antibodies ◽  
Patient Withdrawal ◽  
Life Threatening ◽  
Drug Dependent ◽  
Vancomycin Treatment

Two patients with leukemia experienced profound thrombocytopenia and refractoriness to platelet transfusion during vancomycin treatment. In one patient, withdrawal of drug and administration of platelet transfusions restored platelet counts to near normal levels (approximately 100 x 10(9)/L), however, subsequent challenge with vancomycin due to recurring infection again precipitated severe thrombocytopenia (platelets less than 10 x 10(9)/L) and life- threatening hemorrhagic symptoms. Potent vancomycin-dependent antiplatelet antibodies were detected in the serum of both patients during the refractory period using staphylococcal protein A rosette formation. Employing a monoclonal antibody-antigen capture enzyme- linked immunosorbent assay (ELISA), the patients were found to have vancomycin-dependent IgG antibodies that bound specifically to platelet glycoproteins (GP) IIb and/or IIIa. One of these antibodies failed to react with platelets deficient in GPIIb/IIIa obtained from an individual with Glanzmann's thrombasthenia. These findings provide the first major evidence for drug-dependent antibodies in association with severe thrombocytopenia and refractoriness to platelet transfusion in alloimmunized leukemia patients and, further, provide the first demonstration of vancomycin-dependent antibodies reactive with platelets.

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